¥1,600,000 (Direct Cost : ¥1,600,000)

We newly produced bioresorbable β-chitin sponge and used it as a scaffold for three-dimentional culture of chondrocytes. Β-chitin was obtained from the pens of Loligo squid and the β-chitin sponge was formed into a pillar shape. We produced cartilage-scaffold composites with cartilage-like layer at its surface by culturing β-chitin sponge attached chondrocytes at its surface for 4 weeks. The mean of DNA contents at week 4 established 2.52-fold more than preculture contents. The mean concentration values of chondroitin sulfate and the content of hydroxyproline continued to increase after week 2. The type II collagen and aggrecan genes were both found to be expressed during the experiment. The overall results of the biochemical analysis, along with the histochemical and immunohistochemical findings and RT-PCR analysis, indicate that the cartilage-like layer in the chondrocytes-β-chitin sponge composite was similar to hyaline cartilage. The electron microscopy scanning also reveals that was filled with chondrocytes and abundant extracellular matrix. The β-chitin sponge can be considered biocompatible with chondrocytes, and an adequate scaffold for the three-dimensional chondrocytes culture. Because this technique can produce a pillar-shaped composite, we will be able to pressfit the composites into the articular cartilage defect without covering the periosterium or suturing the implant.